Degree Project in Molecular Biotechnology
Masters Programme in Molecular Biotechnology Engineering, Uppsala University School of Engineering
UPTEC X 15 021 Date of issue 2015-08 Author
Sigrid Lundin
Title (English)
A yeast three-hybrid (Y3H) screen to evolve BirA biotin ligase variants with histone biotinylation activity
Title (Swedish)
Abstract
Histones are heavily modified and marked with epigenetic tags. How these essential proteins together with their modifications are faithfully transmitted through mitotic cell division is still an unanswered question. Such process could be studied using a synthetic modification to tag and trace histone proteins. In vivo lysine biotinylation of histones could ideally serve as a synthetic marker. To be able to bind biotin to histones in vivo, an enzyme with that capacity would be necessary. Preferably an engineered specific enzyme that only biotinylate histones.
BirA is an enzyme that regulates biotinylation in eukaryotic and prokaryotic cells. This project is an attempt to create a BirA mutant through directed evolution, with the capability to biotinylate histones in vivo.
Keywords
Yeast three-hybrid, Y3H, BirA, Biotin, directed evolution.
Supervisors
Simon Elsässer Karolinska Institutet Scientific reviewer
Helena Jernberg Wiklund Uppsala Universitet
Project name Sponsors
Language
English Security
ISSN 1401-2138 Classification
Supplementary bibliographical information Pages
36
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